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Human TNF-alpha ELISA

RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples.

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Antigen Information

Accession Number:
  • P01375
Gene ID:
  • 7124
Gene Symbols:
  • TNF
  • TNFA
  • TNFSF2
Protein Name & Synonyms:
Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Target Species:
  • Human

Assay Format

Detection Range:
30 pg/ml - 6000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
30 pg/ml

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This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4
Number of Targets Detected:
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.


The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Fermoselle C., Rabinovich R., Ausin P., et al. Does oxidative stress modulate limb muscle atrophy in severe COPD patients? Eur Respir J. 2012 Oct;40(4):851-62.
    Species: Human
    Sample type: Tissue Lysate
  2. FilkovÃ? M, HulejovÃ? H, KuncovÃ? K, et al. Resistin in idiopathic inflammatory myopathies. Arthritis Research & Therapy. 2012;14(3):R111. doi:10.1186/ar3836.
    Species: Human
    Sample type: Serum
  3. Rocher C., Singla R., Singal P., et al. Bone morphogenetic protein 7 polarizes THP-1 cells into M2 macrophages. Can J Physiol Pharmacol. 2012 Jul;90(7):947-51. doi: 10.1139/y2012-102.
    Species: Human
    Sample type: Conditioned Media
  4. Sarangi A., Mohapatra P., Dalai R. Serum cytokine TNF-alpha and hemoglobin levels in Plasmodium falciparum malaria - A correlative study in coastal districts of Odisha
    Species: Human
    Sample type: Serum
  5. Siriwardhana N, Layman R, Karwandyar A, Patel S, Tage B, et al. (2012) Inflammatory Cytokines Link Obesity and Breast Cancer. J Metabolic Synd 1:102. doi:10.4172/2167-0943.1000102
    Species: Human
    Sample type: Conditioned Media
  6. Sadik N., Mohamed W., Ahmed M. The association of receptor of advanced glycated end products and inflammatory mediators contributes to endothelial dysfunction in a prospective study of acute kidney injury patients with sepsis. Mol Cell Biochem. 2012 Jan;359(1-2):73-81. doi: 10.1007/s11010-011-1001-4.
    Species: Human
    Sample type: Serum
  7. Tsai H., Chen P., Liu C., et al. Association of hepatitis C virus infection and malnutrition-inflammation complex syndrome in maintenance hemodialysis patients. Nephrol Dial Transplant. 2012 Mar;27(3):1176-83. doi: 10.1093/ndt/gfr458.
    Species: Human
    Sample type: Serum
  8. Goktas C., Cosun A., Bicik Z., et al. Evaluating ESWL-induced renal injury based on urinary TNF-?, IL-1?, and IL-6 levels. Urol Res. 2012 Oct;40(5):569-73.
    Species: Human
    Sample type: Urine
  9. Liou H., Shih C., Chao Y., et al. Inflammatory response to colloids compared to crystalloid priming in cardiac surgery patients with cardiopulmonary bypass. Chin J Physiol. 2012 Jun 30;55(3):210-8. doi: 10.4077/CJP.2012.AMM122.
    Species: Human
    Sample type: Serum
  10. Proksch S., Steinberg T., Keller C., et al. Human saliva exposure modulates bone cell performance in vitro. Clin Oral Investig. 2012 Feb;16(1):69-77. doi: 10.1007/s00784-010-0506-7.
    Species: Human
    Sample type: Conditioned Media
Great kit. Easy to follow protocol. Getting great results.

Florida A&M University
It's so easy to use. I'm happy with it and recommend it to everyone.

Columbia University
Simple protocol and reproducible results. Would recommend.

University of Houston
I used it for human corneal epithelial cell culture supernatant under induced conditions. It is very sensitive and easy to use.

I also want to comment on the fast delivery of the product.

University of Illinois at Chicago
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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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